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PMID: 19790107 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Isolation and partial purification of the Saccharomyces cerevisiae cytokinetic apparatus.

Cytoskeleton (Hoboken, N.J.) ·Vol. 67 ·No. 1 ·2010-01-00 ·Pages 13-22

Young BA, Buser C, Drubin DG

Abstract

Cytokinesis is the process by which a cell physically divides in two at the conclusion of a cell cycle. In animal and fungal cells, this process is mediated by a conserved set of proteins including actin, type II myosin, IQGAP proteins, F-BAR proteins, and the septins. To facilitate biochemical and ultrastructural analysis of cytokinesis, we have isolated and partially purified the Saccharomyces cerevisiae cytokinetic apparatus. The isolated apparatus contains all components of the actomyosin ring for which we tested-actin, myosin heavy and light chain, and IQGAP-as well as septins and the cytokinetic F-BAR protein, Hof1p. We also present evidence indicating that the actomyosin rings associated with isolated cytokinetic apparati may be contractile in vitro, and show preliminary electron microscopic imaging of the cytokinetic apparatus. This first successful isolation of the cytokinetic apparatus from a genetically tractable organism promises to make possible a deeper understanding of cytokinesis.

MeSH Terms
Actomyosin/genetics,isolation & purification,metabolism Cytokinesis/physiology Glucan 1,3-beta-Glucosidase/genetics,isolation & purification,metabolism Green Fluorescent Proteins/genetics,metabolism Myosin Heavy Chains/genetics,isolation & purification,metabolism Recombinant Fusion Proteins/genetics,isolation & purification,metabolism Saccharomyces cerevisiae/metabolism,ultrastructure Saccharomyces cerevisiae Proteins/genetics,isolation & purification,metabolism
Chemicals
MYO1 protein, S cerevisiae Recombinant Fusion Proteins Saccharomyces cerevisiae Proteins Green Fluorescent Proteins Actomyosin Glucan 1,3-beta-Glucosidase Myosin Heavy Chains
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Young Brian A
Department of Molecular and Cell Biology, University of California, Berkeley, California 94720, USA.
Buser Christopher
Drubin David G
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Article Info
Journal
Cytoskeleton (Hoboken, N.J.)
Abbr.
Cytoskeleton (Hoboken)
ISSN
1949-3592
DOI
10.1002/cm.20412
Published
2010-01-00
Pages
13-22
Language
English
Region
United States
NLM ID
101523844
PMCID
PMC2825282
Subset
IM
Grants
NIGMS NIH HHS · R01 GM042759 · United States
NIGMS NIH HHS · R37 GM042759 · United States
NIGMS NIH HHS · R37 GM042759-21 · United States
NIGMS NIH HHS · R01 GM42759 · United States
External Links
PubMed source DOI: 10.1002/cm.20412 PMC Full Text View journal details
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